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The Effects of Alismatis Rhizoma Decoction on the Relative Quantity of Cholesterol Related Genes in Mice

Author: PengMin
Tutor: YuShangGong
School: Hubei University of Chinese Medicine
Course: Pharmaceutical Analysis
Keywords: Alisma soup Hyperlipidemia Lipid-lowering mechanism Synergy Gene Expression
CLC: R285
Type: Master's thesis
Year: 2011
Downloads: 52
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Abstract

Objective: From medical point of view, the pathogenesis of hyperlipidemia is the vacuity of the virtual clinical symptoms of spleen qi deficiency and liver and kidney, the excess of clinical symptoms of blood stasis phlegm. Hyperlipidemia phlegm generation has very close ties with the spleen pathogenesis characteristics in the virtual water transport liquid, wet evil take Qing Yang, causing the patient to suffer from dizziness. So the two homologous different flow, and thus can be used to treat branch drink Alisma soup for the treatment of hyperlipidemia, which actually belong to different diseases, treatment of hyperlipidemia with the party to meet the basic theory of TCM . Modern pharmacology and clinical studies have demonstrated the the Alisma soup has a very significant adjustment dyslipidemia role, have a good effect on hyperlipidemia. The Alisma soup lipid-lowering aspect of the research is still relatively rare, the existing mechanism has been elaborated: inhaling exogenous cholesterol affect the body, or interfere with the body's endogenous cholesterol catabolism, and interfere with the body's anti-fat quality peroxidation. This also reflects the overall treatment effect of multiple targets Alisma soup of lipid metabolism disorders. But from the perspective of molecular biology to study the mechanism of the Alisma soup and lipids, are still in the exploratory stage. High fat diet experiment made the experimental model of hypercholesterolemia mice was measured by observing changes in liver tissue pathological morphology, and serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) content of the number of mouse liver to study high cholesterol features on the extent of damage; the use of the reverse transcription polymerase reaction (RT-PCR) method, determination of the 3 - hydroxy-3 - methyl-glutaryl enzyme A reductase (HMG-CoA reductase) expression in the liver tissue, sterols response element binding protein factor -2 (SREBF-2) 7α hydroxylase (CYP7α1) of cholesterol metabolism gene expression activity in the body, to study the metabolic disorders of the liver lipid substances of high cholesterol molecules mechanism of action. Using ordinary feed conventional adaptation method: select male mice, experimental to keeping 1w mice; 8 after random group 1 (control group) to continue to give a normal diet, given high fat diet for the remaining 40 high-fat made mold, 3w end of the vein blood serum was separated with a semi-automatic biochemical analyzer serum lipids, blood lipid levels is illustrated successful modeling, stochastic modeling success all mice were divided into five groups, each group of eight. The successful modeling 0.01m1 · g-1 dose groups of mice once a day consecutive 4w. (1) blank control group: given ordinary feed, contain saline gavage; (2) the model control group: given ordinary feed, contain saline gavage; (3) the positive control group: given high fat diet at the same time to 6.7 mg · kg-1 simvastatin orally; (4) the Alisma group: given high fat diet to 2g · ml-1 Alisma unilateral solution gavage; (5) Atractylodes group: given high fat diet while 2g · Atractylodes ml-1 the unilateral solution gavage; (6) Compound group (Alisma: the Atractylodes = 5:2): high fat diet at the same time gavage 2g · ml-1 Alisma decoction solution. Mice in each group were reared in separate cages, they can be free to drinking water and food intake. During the experiment is frequently observed in mice state. Gavage daily morning such as above described. Week weighing the teams mice, according to the body weight of each mouse to adjust the dose. Fasting, carotid artery blood serum obtained for the determination of the following indicators: the concentration of serum total cholesterol (TC), triglyceride (TG) concentrations of high-density lipoprotein cholesterol (HDL-C) content, ALT and AST levels . Liver for tissue slices, RNA, and lipid extraction, and organ coefficient was measured at the end of the experiment. 2 liver tissue morphology and pathological changes will be the removal of the liver tissue first photograph, then passes through a formaldehyde-fixed, paraffin-embedded, sliced, then after HE staining by light microscopy to observe the changes in liver histopathology situation. 3 of the total lipids extracted from the liver component first extracted in accordance with the procedure given in the kit instructions, the total lipid in the liver, and then measuring cholesterol in the liver. Serum cholesterol indicators of cholesterol in the liver by the RT-9600 automatic biochemical analyzer to measure these indicators of blood lipids, serum ALT and serum AST. Determination of lipids in the liver tissue of a series of genes mRNA relative expression levels of the two-step method to extract total RNA (β-Actin as a loading control was determined by RT-PCR method, HMG-CoA reductase, SREBF-2 and CYP7α1 mRNAs relative expression level. Results: 1. Alisma, Atractylodes and compound to reduce serum cholesterol indicators role with the model group compared to mouse serum TG and TC concentrations in the the Alisma group and compound groups have emerged to significantly reduce, at the same time serum HDL-C concentrations increased significantly, Alisma, Atractylodes compound mice serum ALT, AST activity increased, with a statistically significant; Alisma, Atractylodes compound could reduce mouse liver, spleen coefficient; ig 4w after, Alisma group compared, the compound can significantly reduce the body weight of mice, and can significantly reduce the liver index. 2. Alisma, Atractylodes and the compound of cholesterol in the liver tissue series gene mRNA relative expression level of When hyperlipidemia, a series of cholesterol metabolism-related genes exhibit abnormal expression of the mouse liver, hyperlipidemia activation of the HMG-CoA reductase accompanied SREBF-2 activation. However, hyperlipidemia mouse liver 7A-hydroxylase gene expression has not been activated. Alisma and combination group showed abnormal expression of cholesterol metabolism genes in our experimental conditions, Alisma and compound decreased serum TC level, while the corresponding mice liver also activate the expression of HMG-CoA reductase. compared with Alismataceae group, the compound also activate the SREBF-2 expression in mouse liver., Alisma and compound mice the liver CYP7α1 gene expression. is not activated. Atractylodes gene expression related to cholesterol metabolism, and does not have a statistically significant compared with the model group, the of Atractylodes mice liver HMG-CoA reductase, CYP7α1 SREBF-2 mRNA of The relative expression quantity no significant change in nature. conclusions: 1. Alisma Decoction reasonable, Alisma, Atractylodes used in combination have a synergistic effect on lowering. 2. Alisma and compound lipid-lowering mechanism may be the HMG-liver CoA reductase gene expression from the down regulation.

The Effects of Alismatis Rhizoma Decoction on the Relative Quantity of Cholesterol Related Genes in Mice

Abstract

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